Use of TP53 reference materials to validate mutations in clinical tissue specimens by single-strand conformational polymorphism analysis
Küçük Resim Yok
Tarih
2004
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
Background: As genetic information moves from basic research laboratories in to the clinical testing environment, there is a critical need for reliable reference materials for the quality assurance of genetic tests. A panel of 12 plasmid clones containing wild-type or point mutations within exons 5-9 have been developed as reference materials for the detection of TP53 mutations. Aim: The goal of this study was to validate the reference materials in providing quality assurance for the detection of TP53 mutations in clinical specimens. Methods: We studied 33 gynecological samples, 11 apparently normal samples and 22 malignant tumors of various origins. Mutations were identified using single-strand conformational polymorphism analysis with both slab gel and capillary electrophoresis. All DNA samples were amplified with fluorescently labeled PCR primers specific for exons 5-9 for mutation detection. Results: Of the 33 patient samples tested, mutations and polymorphisms were found in six specimens in three of the five exons scanned; no mutations were found in exons 7 or 9. Both a mutation and polymorphism were found in non-malignant specimens from the control group. The mutations were confirmed by DNA sequence analysis of the regions scanned. Conclusions: Mutations and polymorphisms were detected in the clinical samples. All of the mutations were silent except for one non-conservative mutation in exon 5, codon 181. This study demonstrates the usefulness of the National Institute of Standards and Technology (NIST) TP53 reference panel in TP53 mutation detection in clinical tissue specimens.
Açıklama
Anahtar Kelimeler
DNA, adult, article, capillary electrophoresis, clinical article, clone, controlled study, DNA sequence, exon, female, fluorescence, gel, gene, gene amplification, gene mutation, genetic polymorphism, human, human tissue, National Institute of Standards and Technology, organization, plasmid, point mutation, polymerase chain reaction, quality control, reference substance, reference value, sequence analysis, single strand conformation polymorphism, tissue, tp53 gene, validation process, Adolescent, Adult, Base Sequence, DNA Mutational Analysis, DNA, Neoplasm, Electrophoresis, Capillary, Exons, Female, Genes, p53, Humans, Middle Aged, Molecular Sequence Data, Mutation, Neoplasms, Polymerase Chain Reaction, Polymorphism, Single-Stranded Conformational, Reference Standards, Sequence Analysis, DNA
Kaynak
Molecular Diagnosis
WoS Q Değeri
Scopus Q Değeri
N/A
Cilt
8
Sayı
2
